Large scale processes are known in the art and are necessary for the industrial production of various materials. Since large scale processes cannot be performed by the same means as small scale processes, specific processes for the large scale production of materials must be designed, even if small scale processes exist.
Nutraceuticals are sometimes prepared using synthetic processes that provide the desired active ingredients, e.g., polyphenols, which are naturally found in fruit cells. However, the use of synthetic processes does not provide the natural ingredients along with the active ingredients, which sometimes contribute to the efficiency of the formulation.
Other types of nutraceuticals are prepared from the natural plants; however, all known large scale processes for preparing nutraceuticals from plants include the extraction of the prepared plant cells in order to obtain the desired active ingredient. However, when plants containing polyphenols, for example, are extracted, the amount of polyphenols, including resveratrol, may be very high in the extraction and therefore, the final product may be bitter. Also, only certain parts of the plant may be successfully extracted since only they contain the desired amounts of the active ingredients.
Small scale processes for the preparation of fruit cells are known in the art; however, large scale processes are more difficult to design since they tend to amplify the production of the primary metabolites, while minimizing the productions of the secondary metabolites. Since active ingredients, such as polyphenols, are secondary metabolites their production in large-scale processes is complex.
Thus, there is a need in the art for a large scale process for preparing fruit cells from natural ingredients, which includes the production of both the primary and the secondary metabolites of the fruit cells.
Metabolic abnormalities are associated with obesity, insulin resistance glucose intolerance, type II diabetes mellitus (DMII), dyslipidemia fatty liver, steatohepatitis, steatosis. These abnormalities increase the risk of stroke cardio-vascular diseases. The etiology of the metabolic syndrome is considered to be multifactorial involving genetic and environmental effects.
Atherosclerosis is a disease of the vascular bed caused by fatty deposit build up in blood vessel walls that narrow the passageway for blood flow within blood vessels. This process may affect all arteries particularly the coronary arteries and may lead to eventual blockage of the coronary arteries resulting in a heart attack, which is the leading cause for premature death in the United States.
Fatty liver disease encompasses a spectrum of clinical conditions characterized histologically mainly by macrovesicular steatosis of the liver. The histopathological spectrum of fatty liver disease ranges from the simple fatty liver (steatosis) to the steatohepatitis, a variant, which has variable degrees of fibrosis. Steatohepatitis may be progressive and can lead to cirrhosis, liver failure and hepatocellular carcinoma and may be a major cause of cryptogenic cirrhosis. The common risk factors for fatty liver disease are obesity, type II diabetes, and hyperlipidemia.
Type II diabetes is among the most common chronic human diseases, affecting almost 8% of the adult population and 19% of people above the age of 65 years in the United States.
Metabolic syndrome also known as “the deadly quartet” or “Syndrome X” or the “Insulin Resistance Syndrome” is a cluster of risk factors for various diseases, such as cardio-vascular diseases, stroke and diabetes mellitus type II, i.e. insulin resistance, hyperinsulinemia, abdominal obesity, (caused by an accumulation of intra-abdominal fat), elevated serum lipids, and high blood pressure. 25% of adults living in the United States are diagnosed with metabolic syndrome. It is believed that the pathophysiology of the metabolic syndrome is related to insulin resistance. The risk factors include the following: elevated waist circumference (≧102 cm in man and 88 cm in women); elevated triglycerides (>150 mg/dL); reduced high-density lipoprotein (HDL) cholesterol (<40 mg/dL in men and 50 mg/dL in women); elevated blood pressure (>130/85 mm Hg) and elevated fasting glucose (>100 mg/dL. Other risk factors may contribute to the metabolic syndrome as well. Additionally, the risk factors may vary in different populations.
Impaired Fasting Glycaemia (IFG) is a pre-diabetic state of dysglycemia associated with insulin resistance, defined as a fasting glucose level of 101-126 mg/dL.
Impaired Glucose Tolerance (IGT) is a pre-diabetic state of dysglycemia associated with insulin resistance defined with similar definition as for IFG or alternatively, defined as a glucose level of 141 to 199 mg/dL two hours following oral consumption of a high glucose meal, typically 75 grams of glucose.
Many of the metabolic maladies are characterized by triglyceride accumulation and insulin resistance.
Triglyceride accumulation in various body tissues, such as muscle, liver and pancreas tissue is considered to be an important factor for organ specific insulin resistance leading to the development of a metabolic malady. Furthermore, accumulation of lipid droplets, which is identical to the term lipid bodies in tissues occurs early in the development of insulin resistance and is correlated with its severity.
Persistent hypertension is one of the risk factors for several diseases and disorders. Hypertension is the single most important modifiable risk factor for ischemic stroke, the third leading cause of death in the Western world. Risk of stroke begins to increase at blood pressure (BP) readings higher than 120/80 millimeters of mercury (mm Hg). Most estimates for hypertension indicate a relative risk of stroke of approximately 4 when hypertension is defined as systolic blood pressure (SBP) ≧160 mm Hg and/or diastolic blood pressure (DSP) ≧95 mm Hg.
Community-based studies have demonstrated that hypertension may contribute to the development of heart failure in as many as 50-60% of patients. In patients with hypertension, the risk of heart failure is increased by 2-fold in men and by 3-fold in women. According to the Framingham Study, hypertension accounts for about one quarter of heart failure cases.
Hypertension is not only a well-established cardiovascular risk factor but also increases the risk of atherosclerosis. Clinical trials have shown that, in the highest quintile of DBP, even with the added risks of high cholesterol and smoking, hypertension still contributes significantly to risk for atherosclerosis.
Hypertension has been reported to occur in 85% to 95% of patients with chronic kidney disease (CKD). Uncontrolled hypertension is a risk factor for developing CKD, and is the second leading cause of end-stage renal disease in the USA.
In the past, most attention was paid to DBP; but nowadays it is recognized that both high SBP and high pulse pressure (the numerical difference between SBP and DBP) are also risk factors. Importantly, it has been demonstrated that the combined evaluation of SBP and DBP improves cardiovascular risk prediction over the 2 individual components. Nevertheless, DBP is still an important cardiovascular risk factor. Isolated diastolic hypertension was a cardiovascular risk factor in the study by Franklin et al. Subjects with isolated diastolic hypertension represented 14% of the hypertensive population, and their cardiovascular risk was found to be about twice that of the subjects with normal BP (Schillaci et al., 2009). Moreover, a nationwide Chinese database also confirmed that isolated diastolic hypertension is an independent risk factor for cardiovascular disease.).
Lipid peroxidation refers to the oxidative deterioration of lipids in a process resulting in cell damage. Lipid peroxidation may be one mechanism through which several risk factors may promote cardiovascular disease.
Lipid peroxidation was found to be highly correlated with cardiovascular risk factors such as age, triglycerides, smoking, cholesterol High-density lipoprotein (HDL) and body mass index (BMI).
Pregnant, oophorectomized, and post-menopausal women exhibit higher levels of lipid peroxidation than non-pregnant, non-oophorectomized and pre-menopausal women, and high level of lipid peroxidation during these states is responsible, at least in part, for their increased risk of CVD.
Low density lipoprotein (LDL) isolated from patients with essential hypertension exhibits increased propensity for oxidation. Thus, oxidized LDL (Ox-LDL) may predispose to accelerated atherosclerosis, and the propensity of LDL for lipid peroxidation may be a risk factor for atherogenesis as important as the plasma concentration of LDL itself.
Flow-mediated dilation (FMD) as an expression of endothelial dysfunction may serve as an indicator of several diseases. FMD was highly predictive for coronary artery disease (CAD) with an odds ratio of 1.32 for each percent decrease in FMD (p=0.001). Not only in atherosclerosis but also in systemic sclerosis FMD was highly correlated with the progression of the disease. There was an inverse correlation between FMD values and disease duration.
In post-menopausal women low FMD values were correlated with an increase in relative risk to developing hypertension.
FMD is associated with increased mortality risk in ischaemic advanced chronic heart failure (ACHF) patients. (Shechter et al., 2009). Moreover, persistent low FMD despite therapies for heart failure (HF) and atherosclerotic risk factors was a predictor of cardiac events in patients with chronic ischemic HF.
Stroke patients had significantly lower flow-mediated dilation (FMD) values than nonstroke patients. FMD as an independent predictor of cryptogenic stroke when adjusted by age, sex, and presence of patent foramen ovale (PFO).
Finally, in children with heterozygous familial hypercholesterolemia (heFH) who are prone to premature atherosclerosis decreased FMD is detected before structural atherosclerotic changes occur.
Nutraceuticals derived from polyphenol-containing fruit extracts are known for their beneficial effects. The use of red wine as a source of these regulatory constituents is limited due to its high alcoholic content and sugar. In addition, it has been shown that the therapeutic effect of wine and wine grapes is dependant on species, location, year (annual climate), processing etc. and therefore reliance on red wine, grapes or grape seeds as a source of these regulatory compounds does not lead to a homogeneous or consistent supply of material. Furthermore, fruits are often contaminated by residual fungicides, pathogens, pesticides and pollutants.
Moreover, the potential benefit of gastrointestinal delivery of polyphenols from red wines and fruit extracts is limited by its bioavailability to target tissues and cells. Due to marked differences in their bioavailability while passing through the intestines, no correlation can be drawn between the abundance of a certain polyphenol in a given food and its concentration as an active compound in vivo. Absorbance of flavonoids in the small intestines, for example, ranges from 0-60% of the dose, and elimination half-lives range from 2-48 hours. Most polyphenols undergo extensive metabolism in the intestine, and are present in serum and urine predominantly as glucuronides, methyl or sulfate. Among the known polyphenols, the phytoalexin resveratrol (trans-3,5,4′-trihydroxystilbene) (RES), found in red grapes, red wine and other foods such as different kind of berries and peanuts has drawn most of the attention. It is believed to be responsible for the “French paradox”, a phenomenon associated with low incidence of cardiovascular diseases despite high-fat diet as a result of moderate red wine consumption. However, RES bioavailability is compromised by its physicochemical properties such as low water solubility and also its high hepatic uptake. Moreover, oral bioavailability of RES is extremely low due to rapid and extensive metabolism with the consequent formation of various metabolites.
Studies investigating RES activity and effects rely mainly on three sources of resveratrol, namely pure synthetic RES, natural plant-derived RES (e.g. Poligonum cupcidatum extracts) products, or to a lesser extent whole red grapes or their products (red wine, grape juice, grape extracts). Red Grape Cells (RGC; Fruitura Bioscience Ltd, Israel) is a natural patent-protected formulation of cultured cells originated from the fruits of Vitis Vinifera L. cultivar comprising the whole matrix of polyphenols including resveratrol and other ingredients naturally existing in red grape.
There is thus a need for natural (phyto) compositions that may be prepared in a large scale process in which the amount of the active ingredient is consistent and recurrent (e.g., clonal preparations), is highly bioavailable and easily administered for the treatment of various disorders, including conditions or complication associated with metabolic disorder or metabolic maladies.